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. Author manuscript; available in PMC: 2010 Jan 1.

Published in final edited form as: Nat Struct Mol Biol. 2009 Jun 14;16(7):731–739. doi: 10.1038/nsmb.1625

Fig. 4 — The interaction of Brr2 with RNA and the effect of Prp8-CTR on these interactions. (a) EMSA demonstrates that Hel308-II does not bind a 13nt ss or dsRNA with arbitrary sequence (GAGAUUUAUUUCG) in the absence or presence of ATP. UAP56, another DExD/H-box protein involved in splicing, is used as a positive control and binds both ss and dsRNA in the presence of ATP. (b) Hel308 (lanes 1–4) and full-length Brr2 (lanes 5–10) do not bind U4/U6 at indicated concentrations but the presence of Prp8-CTR generate a complex that binds U4/U6 better. Supershift experiments using anti-Brr2 and anti-Prp8 antibodies confirm the identity of the shifted bands (lanes 11–13). (c) Titration experiments indicate that Prp8-CTR binds U4/U6 with a Kd of 2.2±0.2 µM. Error bars and errors in Kd are s.d. d) Prp8-CTR binds ss or dsR13 very weakly.

Fig. 4 — The interaction of Brr2 with RNA and the effect of Prp8-CTR on these interactions. (a) EMSA demonstrates that Hel308-II does not bind a 13nt ss or dsRNA with arbitrary sequence (GAGAUUUAUUUCG) in the absence or presence of ATP. UAP56, another DExD/H-box protein involved in splicing, is used as a positive control and binds both ss and dsRNA in the presence of ATP. (b) Hel308 (lanes 1–4) and full-length Brr2 (lanes 5–10) do not bind U4/U6 at indicated concentrations but the presence of Prp8-CTR generate a complex that binds U4/U6 better. Supershift experiments using anti-Brr2 and anti-Prp8 antibodies confirm the identity of the shifted bands (lanes 11–13). (c) Titration experiments indicate that Prp8-CTR binds U4/U6 with a Kd of 2.2±0.2 µM. Error bars and errors in Kd are s.d. d) Prp8-CTR binds ss or dsR13 very weakly.